黄芩苷对脂多糖诱导的小鼠炎症的保护作用
Effects of baicalin on protecting inflammation in mice induced by lipopolysaccharide
闫普普 1朱君 1刘佳丽 1黄永熙 1余捷 1汤锋 1郭利伟1
作者信息
- 1. 长江大学动物科学技术学院,荆州 434000
- 折叠
摘要
为进一步明晰黄芩苷(BCN)对脂多糖(LPS)诱导的小鼠炎症损伤的缓解作用机制,采用体内体外试验结合,体外试验采用LPS建立RAW264.7细胞炎症模型,测定细胞吞噬能力、一氧化氮释放、炎症细胞因子及TGF-β/SMAD2通路相关mRNA表达;体内试验采用不同浓度的黄芩苷对BALB/c小鼠连续7 d灌胃后腹腔注射LPS建立炎症模型,测定小鼠脾脏指数、T细胞亚群及免疫平衡,RT-qPCR法测定脾脏炎症细胞因子和TGF-β/SMAD2通路相关基因的mRNA表达.结果显示:黄芩苷在0～25 μg/mL范围内对RAW264.7增殖无抑制作用,LPS质量浓度为1 mg/mL时,细胞存活率为54.55%.不同质量浓度黄芩苷均可增强RAW264.7细胞的吞噬能力(P<0.05),降低NO释放(P<0.05);LPS刺激后炎症细胞因子mRNA表达上升(P<0.05),TGF-β和SMAD2表达下降,黄芩苷干预后上述mRNA表达得到逆转.此外,LPS处理后小鼠脾脏指数显著上升(P<0.05),不同剂量的黄芩苷组均改善这一情况(P<0.05).黄芩苷能够改善LPS刺激的CD4+细胞与CD8+细胞的分化,降低CD4+/CD8+,同时,黄芩苷可恢复LPS刺激的Th17/Treg平衡轴失衡.结果表明,黄芩苷对LPS诱导的小鼠炎症具有缓解作用,其作用机制可能与TGF-β/SMAD2信号通路激活、抑制炎症因子的表达及恢复Th17/Treg平衡轴有关.
Abstract
A inflammatory model of RAW264.7 cell induced with lipopolysaccharide(LPS)was es-tablished in vitro to study the effects and mechanism of baicalin(BCN)against LPS-induced inflammato-ry injury in mice.The cellular phagocytic ability,nitric oxide(NO)release,and mRNA expression to in-flammatory cytokines and TGF-β/SMAD2 pathways related genes were measured.Experiments in vivo were conducted using different concentrations of baicalin to establish an inflammatory model in BALB/c mice after 7 consecutive days of gastric lavage and intraperitoneal injection of LPS.The spleen index,sub-groups and immune balance of T cell in mice were detected.RT-qPCR was used to determine the mRNA expression of inflammatory cytokines and TGF-β/SMAD2 pathways related genes in the spleen.Results showed that baicalin had no effect on inhibiting the proliferation of RAW264.7 cell within the range of 0-25 μg/mL.When the LPS concentration was 1 mg/mL,the survival rate of RAW264.7 cell was 54.55%.Different concentrations of baicalin enhanced the phagocytic ability(P<0.05)and reduced NO release(P<0.05)of RAW264.7 cells.The mRNA expression of inflammatory cytokines related genes increased(P<0.05)while the mRNA expression of TGF-β/SMAD2 pathways related genes decreased after LPS stimulation.The mRNA expression of above genes was reversed after intervention with baicalin.The spleen index of mice significantly increased(P<0.05)after LPS treatment,and different doses of baicalin groups improved this situation(P<0.05).Results of flow cytometry showed that baicalin improved the dif-ferentiation of CD4+and CD8+cells stimulated by LPS,reduced the ratio of CD4+/CD8+,and restored the imbalance of Th17/Treg balance axis induced by LPS.It is indicated that baicalin alleviates the LPS induced inflammation in mice,and its mechanism may be related to the activation of TGF-β/SMAD2 sig-naling pathway,the inhibition of inflammatory cytokines expression,and the restoration of Th17/Treg bal-ance axis.
关键词
黄芩苷/脂多糖/免疫调节/TGF-β/SMAD2/T细胞分化Key words
baicalin(BCN)/lipopolysaccharide(LPS)/immune regulation/TGF-β/SMAD2/dif-ferentiation of T cell引用本文复制引用
出版年
2024
NETL
NSTL
万方数据